Data fusion between high resolution (1)H-NMR and mass spectrometry: a synergetic approach to honey botanical origin characterization.

A data fusion approach was applied to a commercial honey data set analysed by (1)H-nuclear magnetic resonance (NMR) 400 MHz and liquid chromatography-high resolution mass spectrometry (LC-HRMS). The latter was performed using two types of mass spectrometers: an Orbitrap-MS and a time of flight (TOF)-MS. Fifty-six honey samples from four monofloral origins (acacia, orange blossom, lavender and eucalyptus) and multifloral sources from various geographical origins were analysed using the three instruments. The discriminating power of the results was examined by PCA first considering each technique separately, and then combining NMR and LC-HRMS together with or without variable selection. It was shown that the discriminating potential is increased through the data fusion, allowing for a better separation of eucalyptus, orange blossom and lavender. The NMR-Orbitrap-MS and NMR-TOF-MS mid-level fusion models with variable selection were preferred as a good discrimination was obtained with no misclassification observed for the latter. This study opens the path to new comprehensive food profiling approaches combining more than one technique in order to benefit from the advantages of several technologies. Graphical Abstract Data fusion between high resolution 1H-NMR and mass spectrometry.

[Sleep, an issue for nurses and patients]. / Le sommeil, un enjeu pour les soignants et les patients.

A three-day training programme on sleep in hospitals was created in 2008 on the initiative of the national association for the continuing training of hospital staff, in collaboration with the French ministry of health. 448 nurses have been trained thanks to a participative programme integrating new, interactive pedagogical tools.

The GMO90+ Project: Absence of Evidence for Biologically Meaningful Effects of Genetically Modified Maize-based Diets on Wistar Rats After 6-Months Feeding Comparative Trial.

The GMO90+ project was designed to identify biomarkers of exposure or health effects in Wistar Han RCC rats exposed in their diet to 2 genetically modified plants (GMP) and assess additional information with the use of metabolomic and transcriptomic techniques. Rats were fed for 6-months with 8 maize-based diets at 33% that comprised either MON810 (11% and 33%) or NK603 grains (11% and 33% with or without glyphosate treatment) or their corresponding near-isogenic controls. Extensive chemical and targeted analyses undertaken to assess each diet demonstrated that they could be used for the feeding trial. Rats were necropsied after 3 and 6 months. Based on the Organization for Economic Cooperation and Development test guideline 408, the parameters tested showed a limited number of significant differences in pairwise comparisons, very few concerning GMP versus non-GMP. In such cases, no biological relevance could be established owing to the absence of difference in biologically linked variables, dose-response effects, or clinical disorders. No alteration of the reproduction function and kidney physiology was found. Metabolomics analyses on fluids (blood, urine) were performed after 3, 4.5, and 6 months. Transcriptomics analyses on organs (liver, kidney) were performed after 3 and 6 months. Again, among the significant differences in pairwise comparisons, no GMP effect was observed in contrast to that of maize variety and culture site. Indeed, based on transcriptomic and metabolomic data, we could differentiate MON- to NK-based diets. In conclusion, using this experimental design, no biomarkers of adverse health effect could be attributed to the consumption of GMP diets in comparison with the consumption of their near-isogenic non-GMP controls.

[Objective improvement of sleep disorders in the elderly by a health education program]. / Les troubles du sommeil peuvent être objectivement améliorés chez les seniors par un programme personnalisé d'éducation à la santé

INTRODUCTION: The prevalence of sleep disorders increases with age and reaches 20 to 40% of those older than 60 years. We set up a health education program to help the elderly to improve their sleep. It includes a preliminary 9-day evaluation with a sleep diary and wrist actigraph, a day of group cognitive behavioral therapy, and a follow-up assessment, again with sleep diary and actigraph. METHODS: Of the 26 study participants (9 men and 17 women, mean age: 68+/-1 years), 14 had insomnia with night awakenings of 1 hour or longer or a sleep latency of 30 minutes or longer or both (group 1). The other 12 (group 2) also complained of insufficient sleep. RESULTS: In the weeks following cognitive behavioral therapy, group 1 improved their total sleep time by an average of 24 to 33 minutes, with reduced night-time awakenings and sleep latency and no change in their time spent in bed. Those in group 2 also increased their total sleep time by 18 to 47 minutes, by spending more time in bed and maintaining a sleep efficiency close to 88%. CONCLUSION: This study showed that cognitive behavioral therapy coupled with individual sleep evaluation improves sleep duration in elderly people who complain of insufficient sleep. These beneficial effects were accompanied by positive assessments of both subjective sleep quality and morning energy.

Development and validation of a multiresidue method for the analysis of more than 500 pesticides and drugs in water based on on-line and liquid chromatography coupled to high resolution mass spectrometry.

Screening of a large number of emerging pollutants is highly desirable for the control of water quality. In this respect, a novel, fully automated contaminant screening method based on an integrated sample preconcentration and liquid chromatography coupled to high resolution mass spectrometry (SPE-UHPLC-HRMS) has been developed. The optimal chromatographic column and experimental conditions allowing the retention and subsequent elution of the maximum number of analytes were defined. Liquid chromatography and Q-exactive (Orbitrap™) parameters were optimized to obtain the best separation of molecules of interest, and the lowest detection limits. Due to the large amount of data to compare, a script written in R language was developed to evaluate the quality of the data generated by the comparison of 14 experimental conditions. The developed method enables the simultaneous semi quantitative analysis of 539 compounds (pesticides and drug residues), in 36 min with only 5 mL of water. Method validation was achieved through studies of repeatability, selectivity, linearity and matrix effect. Application to 20 tap water samples collected in and around Paris showed the presence of 34 different compounds all with concentrations below 0.1 µg/L, the European Union limit for drinking water. Pesticides and transformation products frequently found in water resources such as atrazine and its metabolites, hexazinone, oxadixyl, propazine and simazine were detected. Drug residues such as valsartan and carbamazepine, usually not monitored, were also found. The next step will be to assess the ability of this method to highlight the presence of unexpected contaminants not present in our database.

Minimum data set for nutritional intervention studies in elderly people.

Malnutrition, considered for the purpose of the present data set as undernutrition, is a major risk factor of mortality in elderly people. Such protein-energy malnutrition should be detected as soon as possible. Once established, this malnutrition state must be corrected by appropriate diet, supplementation, artificial nutrition, or therapeutic treatment. If carried out well, these interventions should reduce the risk of mortality and, for some diseases such as degenerative diseases, may postpone morbidity and dependence. The efficiency of nutritional interventions has already been evaluated by different means including the measurement of anthropometric and laboratory parameters. However, in the absence of a consensus on the use of these parameters, comparison between studies and even effectiveness of the proposed treatment are frequently unconvincing. The relevance of the most common markers used in epidemiologic studies on malnutrition and nutritional interventions in elderly persons was studied for establishing a minimum data set. The aim of this task force was to provide investigators and operators in the field of clinical nutrition with clear and expert validated clinical outcomes allowing them to design and set up conclusive trials.

[Sleep disorders in elderly]. / Troubles du sommeil chez le sujet agé.

Insomnia affects 20% of the adult population in western countries and its prevalence increases with age. There is a controversy regarding the origin of sleep disorders in elderly. Are they only due to a senile process of sleep functioning or due to other associated comorbidities? Considering the objective assessment of sleep in elderly (by polysomnography), it has been shown an increasing sleep latency, decreasing total sleep time and sleep efficiency, a lower percentage of slow wave sleep. The circadian clock is also modified by age with phase advance and a decreased amplitude of the circadian rhythms. The most relevant comorbidities found in older people are: sleep apneas, restless leg syndrome, psychiatric disorders (anxiety and depression) and the use of drugs.

High-resolution mass spectrometry associated with data mining tools for the detection of pollutants and chemical characterization of honey samples.

Analytical methods for food control are mainly focused on restricted lists of well-known contaminants. This paper shows that liquid chromatography-high-resolution mass spectrometry (LC/ESI-HRMS) associated with the data mining tools developed for metabolomics can address this issue by enabling (i) targeted analyses of pollutants, (ii) detection of untargeted and unknown xenobiotics, and (iii) detection of metabolites useful for the characterization of food matrices. A proof-of-concept study was performed on 76 honey samples. Targeted analysis indicated that 35 of 83 targeted molecules were detected in the 76 honey samples at concentrations below regulatory limits. Furthermore, untargeted metabolomic-like analyses highlighted 12 chlorinated xenobiotics, 1 of which was detected in lavender honey samples and identified as 2,6-dichlorobenzamide, a metabolite of dichlobenil, a pesticide banned in France since 2010. Lastly, multivariate statistical analyses discriminated honey samples according to their floral origin, and six discriminating metabolites were characterized thanks to the MS/MS experiments.

Relevance of the cyclophosphamide-induced cystitis model for pharmacological studies targeting inflammation and pain of the bladder.

This work aimed at establishing the relevance of using the in vivo model of cyclophosphamide (CYP)-induced bladder inflammation in rats for in vivo pharmacological studies. Specifically, we measured visceral nociception, identified key inflammatory mediators and evaluated the effects of relevant pharmacological treatments. Cystitis was induced in female rats by a single CYP injection. Sensitivity of the lower abdomen to von Frey mechanical stimulation was determined as a nociceptive parameter. Bladders were assessed for weight, wall thickness and macroscopic damage. Inflammatory mediators were quantified in bladders and urines. The effects of aspirin, ibuprofen and morphine were investigated on all these parameters. A single CYP injection increased nociceptive scores and decreased nociceptive threshold in response to mechanical stimuli between 1 and 4h post-administration. Increased bladder weight and wall thickness were associated with edema and hemorrhage. Bladder levels of IL-1ß, IL-6, MCP-1 and VCAM, and urinary levels of PGE2 were increased. In contrast, a decrease in the urinary metabolites, indoxyl sulfate and pantothenic acid, was observed. Aspirin, ibuprofen and morphine decreased CYP-induced referred visceral pain. Aspirin and ibuprofen also reversed the increased wall thickness, macroscopic damage and levels of IL-1ß, IL-6 and PGE2, and the decreased panthotenic acid levels. In contrast, morphine increased wall thickness, edema, hemorrhage, and bladder IL-6 and MCP-1 levels. This work presents a new and reliable method to evaluate visceral sensitivity in rats, and new relevant biomarkers identified in the bladder and urine to measure inflammation and pain parameters for in vivo pharmacological studies.

[Renal aging: risk factors and nephroprotection]. / Vieillissement rénal: facteurs de risque et néphroprotection.

Renal failure in the elderly is currently underestimated and presents a real challenge for the public health system. Kidney function must be routinely assessed by creatinine clearance, estimated with either the Cockcroft and Gault formula or the simplified MDRD formula, which appears especially appropriate for the elderly. Normal kidney aging is related to tissue and functional changes that make older patients very vulnerable to environmental modifications. Numerous factors can accelerate the impairment of rental function during aging. Some of them cannot be modified: sex, genome, and initial kidney disease. Most of them can be managed or treated: hypertension, diabetes mellitus, obesity, smoking, dyslipidemia, proteinuria, and the presence of oxidation and glycation products. Chronic renal failure in the elderly must be managed early with strict treatment targets to avoid the development of end-stage renal disease. Inhibitors of the renin-angiotensin-aldosterone system play an essential role in optimizing nephroprotection: control of hypertension, diabetes complications, and proteinuria. They should be prescribed very carefully in older patients. Age is not a prerequisite for consultations with nephrologists, which should take place early so that nephroprotection can still be useful.

Nutritional assessment of residents in long-term care facilities (LTCFs): recommendations of the task force on nutrition and ageing of the IAGG European region and the IANA.

Unintentional weight loss and Undernutrition are major problems among older people living in Long-Term Care Facilities (LTCF). Undernutrition manifests in LTCF particularly as weight loss and low Body Mass Index (BMI) and is associated with increased morbidity and mortality as well as with functional decline. There are many factors associated with poor nutritional status and affecting protein-energy intake and/or energy expenditure. These include age of 85 years or older, low nutrient intake, loss of ability to eat independently, swallowing and chewing difficulties, becoming bed-ridden, pressure ulcers, history of hip fracture, dementia, depressive symptoms and suffering from two or more chronic illnesses. Nutritional evaluation is an essential part of the Comprehensive Geriatric Assessment (CGA). This evaluation ranges from methods such as BMI to several validated tools such as Mini-Nutritional Assessment (MNA). After diagnosis, the management of undernutrition in LTCF requires a multidisciplinary approach which may involve dietary and environmental improvements and managing multiple co-morbidities, while avoiding polypharmacy as far as possible. Finally, the need for supplementation or artificial (tube) feeding may be considered taking into account the CGA and individual needs. This document presents a succinct review and recommendations of evaluation and treatment of undernutrition.

Low-dose aspirin prevents age-related endothelial dysfunction in a mouse model of physiological aging.

The age-related impairment of endothelium-dependent vasodilatation contributes to increased cardiovascular risk in the elderly. For primary and secondary prevention, aspirin can reduce the incidence of cardiovascular events in this patient population. The present work evaluated the effect of low-dose aspirin on age-related endothelial dysfunction in C57B/J6 aging mice and investigated its protective antioxidative effect. Age-related endothelial dysfunction was assessed by the response to acetylcholine of phenylephrine-induced precontracted aortic segments isolated from 12-, 36-, 60-, and 84-wk-old mice. The effect of low-dose aspirin was examined in mice presenting a decrease in endothelial-dependent relaxation (EDR). The effects of age and aspirin treatment on structural changes were determined in mouse aortic sections. The effect of aspirin on the oxidative stress markers malondialdehyde and 8-hydroxy-2'-deoxyguanosine (8-OhdG) was also quantified. Compared with that of 12-wk-old mice, the EDR was significantly reduced in 60- and 84-wk-old mice (P < 0.05); 68-wk-old mice treated with aspirin displayed a higher EDR compared with control mice of the same age (83.9 +/- 4 vs. 66.3 +/- 5%; P < 0.05). Aspirin treatment decreased 8-OHdG levels (P < 0.05), but no significant effect on intima/media thickness ratio was observed. The protective effect of aspirin was not observed when treatment was initiated in older mice (96 wk of age). It was found that low-dose aspirin is able to prevent age-related endothelial dysfunction in aging mice. However, the absence of this effect in the older age groups demonstrates that treatment should be initiated early on. The underlying mechanism may involve the protective effect of aspirin against oxidative stress.

Changes in rat liver mitochondria with aging. Lon protease-like reactivity and N(epsilon)-carboxymethyllysine accumulation in the matrix.

Aging is accompanied by a gradual deterioration of cell functions. Mitochondrial dysfunction and accumulation of protein damage have been proposed to contribute to this process. The present study was carried out to examine the effects of aging in mitochondrial matrix isolated from rat liver. The activity of Lon protease, an enzyme implicated in the degradation of abnormal matrix proteins, was measured and the accumulation of oxidation and glycoxidation (Nepsilon-carboxymethyllysine, CML) products was monitored using immunochemical assays. The function of isolated mitochondria was assessed by measuring respiratory chain activity. Mitochondria from aged (27 months) rats exhibited the same rate of oxygen consumption as those from adult (10 months) rats without any change in coupling efficiency. At the same time, the ATP-stimulated Lon protease activity, measured as fluorescent peptides released, markedly decreased from 10-month-old rats (1.15 +/- 0.15 FU x micro g protein-1 x h-1) to 27-month-old-rats (0.59 +/- 0.08 FU x micro g protein-1 x h-1). In parallel with this decrease in activity, oxidized proteins accumulated in the matrix upon aging while the CML-modified protein content assessed by ELISA significantly increased by 52% from 10 months (11.71 +/- 0.61 pmol CML x micro g protein-1) to 27 months (17.81 +/- 1.83 pmol CML x micro g protein-1). These results indicate that the accumulation of deleterious oxidized and carboxymethylated proteins in the matrix concomitant with loss of the Lon protease activity may affect the ability of aging mitochondria to respond to additional stress.

Gene expression in aging kidney and pituitary.

Gene expression in aging kidney and pituitary was determined by subtractive hybridization, DNA microarrays and RT-PCR. Kidneys and pituitary were removed from 10- and 30-month-old female WAG/Rij rats, which were free from chronic progressive nephrosis and had a low incidence of pituitary tumors with age. From 350 cDNA fragments isolated by subtractive hybridization, just one showed a more than twofold change in expression between 10 and 30 months. The use of a specific microarray with 4050 rodent genes also failed to detect downregulation lower than 0.5 or upregulation larger than 2.0 in aging rat kidney. Similarly, mRNA content for vasopressin V2 and V1 receptors, aquaporin 2 and 3, and adenylyl cyclase type VI was not significantly modified with age as determined by RT-PCR. In contrast, microarray analysis of pituitary mRNA expression showed upregulation of 11 genes with ratios equal to or greater than 2.0 and downregulation of 6 genes with ratios equal to or less than 0.5. Two cDNA sequences of unknown genes from the kidney subtractive library were part of the age-related up- and downregulated genes of the pituitary. Other genes were mainly related to cell differentiation, control of homeostasis, cellular signaling, endoplasmic reticulum trafficking and metabolism. These data indicated that mRNA expression is barely modified in aging kidney free from chronic progressive nephrosis, at least in the 0.5-2.0 range, in contrast to pituitary. They also suggest that the downregulation of proteins reported in aging kidneys free from gross disease is related to post-transcriptional changes.

Food restriction prevents advanced glycation end product accumulation and retards kidney aging in lean rats.

ABSTRACT.: Tissue content of advanced glycation end products (AGE) increases with age and contributes to the changes in structure and function of the renal and cardiovascular systems. The effect of chronic food restriction on this AGE accumulation was investigated in lean WAG/Rij rats. A 30% food restriction performed from 10 to 30 mo in female rats reduced their mean body weight from 240 +/- 7 to 160 +/- 12 g, but did not modify their survival. AGE collagen content increased from 14.3 +/- 5.5 to 104.7 +/- 13.0 arbitrary units per microgram (AU/microg) of hydroxyproline (OHPro) in kidney between 10 and 30 mo, and from 9.7 +/- 1.2 to 310.6 +/- 34.6 AU/microg OHPro in the abdominal aorta. Food restriction reduced AGE accumulation to 21.4 +/- 3.3 and 74.6 +/- 16.5 AU/microg OHPro in kidney and aorta of 30-mo-old animals. Similar results were found for collagen prepared from isolated glomeruli (7.8 +/- 1.2, 81.2 +/- 16.1, and 10.3 +/- 4.3 AU/microg OHPro in 10-mo, 30-mo, and restricted 30-mo-old rats). Reduction of intrarenal and arterial AGE accumulation by food restriction was confirmed by immunostaining in optical microscopy. Age-related changes in arterial and kidney structures as polyuria and proteinuria were mainly prevented by food restriction. These data indicate that chronic food restriction reduces the accumulation of AGE and preserves the structure and function of the renal and cardiovascular systems in learn rats, although it did not affect survival of the animals between 10 and 30 mo.

Correction of age-related polyuria by dDAVP: molecular analysis of aquaporins and urea transporters.

Senescent female WAG/Rij rats exhibit polyuria without obvious renal disease or defects in vasopressin plasma level or V(2) receptor mRNA expression. Normalization of urine flow rate by 1-desamino-8-d-arginine vasopressin (dDAVP) was investigated in these animals. Long-term dDAVP infusion into 30-mo-old rats reduced urine flow rate and increased urine osmolality to levels comparable to those in control 10-mo-old rats. The maximal urine osmolality in aging rat kidney was, however, lower than that in adult kidney, despite supramaximal administration of dDAVP. This improvement involved increased inner medullary osmolality and urea sequestration. This may result from upregulation of UT-A1, the vasopressin-regulated urea transporter, in initial inner medullary collecting duct (IMCD), but not in terminal IMCD, where UT-A1 remained low. Expression of UT-A2, which contributes to medullary urea recycling, was greatly increased. Regulation of IMCD aquaporin (AQP)-2 (AQP2) expression by dDAVP differed between adult and senescent rats: the low AQP2 abundance in senescent rats was normalized by dDAVP infusion, which also improved targeting of the channel; in adult rats, AQP2 expression was unaltered, suggesting that IMCD AQP2 expression is not regulated by dDAVP directly. Increased AQP3 expression in senescent rats may also be involved in improved urine-concentrating capacity owing to higher basolateral water and urea reabsorption capacity.